Culture medium and device for detection and enumeration of enterobacteriaceae

• US 5,601,998 A
• Filed: 08/18/1994
• Issued: 02/11/1997
• Est. Priority Date: 08/18/1994
• Status: Expired due to Term

• Current Assignee: 3M Company
• Original Assignee: 3M Company
• Inventors: Mach, Patrick A., Wickert, Peter D., Adams, Carl A.
• Primary Examiner(s): Kight, John
• Assistant Examiner(s): Leary, Louise N.
• Application Number:
  US08/292,784
• Time in Patent Office: 908 Days
• Field of Search: 435/34, 435/4, 435/29, 435/12, 435/805, 435/23, 435/848, 435/849, 435/38, 436/501, 436/74, 436/63, 422/55, 422/57, 422/68.1
• US Class Current: 435/34
• CPC Class Codes: C12M 23/04 Flat or tray type, drawers C12M23/10, C12M23/12, C12M23/16 take precedence
• CPC Class Codes: C12M 23/22 Transparent or translucent parts glassware for laboratory use B01L3/00
• CPC Class Codes: C12M 23/26 flexible flexible containers for laboratory use B01L3/505
• CPC Class Codes: C12M 23/38 Caps; Covers; Plugs; Pouring means
• CPC Class Codes: C12M 25/06 Plates; Walls; Drawers; Multilayer plates
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12Q 1/045 Culture media therefor
• CPC Class Codes: C12Q 1/10 Enterobacteria
• CPC Class Codes: Y10S 435/848 Escherichia
• CPC Class Codes: Y10S 435/849 Escherichia coli

Abstract

This invention generally relates to products and processes used to determine the presence of Enterobacteriaceae in a sample and particularly relates to a bacterial culture medium which may be used in products and processes to allow early detection and enumeration of Enterobacteriaceae in a sample. The bacterial culture medium which facilitates the early detection and enumeration of Enterobacteriaceae contains a selected amount of glucose, pH indicator and buffer which prevent diffusion of colored indicator zones associated with growing bacteria in the medium.

Claims

1. A bacterial culture medium comprising glucose and a pH indicatorbuffer system to detect and enumerate EntCrobacteriaceae growing in the medium wherein the medium contains selected amounts of glucose, pH indicator and buffer to prevent colored indicator zones from excessively diffusing in the medium.

2. The culture medium of claim 1 wherein a sample containing Enterobacteriaceae is added to the culture medium in an aqueous diluent consisting essentially of about 0.6 mMol sodium phosphate.

3. The culture medium of claim 2 comprising about 7-14 g/l gelatin peptone, 6-9 g/l yeast extract, 5-20 g/l glucose, 5-15 g/l sodium chloride, 2.7-3.15 g/l bile salts, 5-13 g/l guar gum, 0.6-2 g/l monobasic potassium phosphate, 1.8-6 g/l dibasic potassium phosphate, and 0.2-0.8 g/l pH indicator.

4. The culture medium of claim 2 containing about 14 gl gelatin peptone, 6 g/l yeast extract, 20 g/l glucose, 10 g/l sodium chloride, 3 g/l bile salts, 11 g/l guar gum, 2 g/l monobasic potassium phosphate, 6 g/l dibasic potassium phosphate, and 0.4 g/l pH indicator.

5. The culture medium of claim 1 wherein a sample containing Enterobacteriaceae is added to the culture medium in an aqueous diluent consisting essentially of about 0.1 wt/vol % casein peptone and 0.85 wt/vol % sodium chloride.

6. The culture medium of claim 5 comprising about 7-14 g/l gelatin peptone, 6-9 g/l yeast extract, 5-20 g/l glucose, 2.7-3.15 g/l bile salts, 5-13 g/l guar gum, 0.2-1 g/l monobasic potassium phosphate, 0.6-3 g/l dibasic potassium phosphate, and 0.2-0.8 g/l pH indicator.

7. The culture medium of claim 5 containing about 14 g/l gelatin peptone, 6 g/l yeast extract, 20 g/l glucose, 3 g/l bile salts, 11 g/l guar gum, 0.4 g/l monobasic potassium phosphate, 1.2 g/l dibasic potassium phosphate, and 0.4 g/l pH indicator.

8. The culture medium of claim 1 wherein pH indicator is selected from the group consisting of azobenzene, sulfonphthalein and anthroquinone pH indicators.

9. The culture medium of claim 8 wherein the pH indicator is selected from the group consisting of methyl red, bromocresol purple, chlorophenol red, bromthymol blue, bromcresol blue, and 3,4-dihydroxy-9, 10-dioxo-2-anthracenesulfonic acid, sodium salt.

10. The culture medium of claim 9 wherein the pH indicator is chlorophenol red.

11. A method of detecting and enumerating Enterobactriaceae in a sample comprising the steps of adding an aliquot of a sample in an aqueous diluent to a culture medium containing selected amounts of glucose, pH indicator and buffer to prevent colored indicator zones from excessively diffusing in the medium, adjusting the pH of the aliquot to about 6.57-7.5;

growing Enterobacteriaceae in the presence of the culture medium; and

detecting the color change of the pH indicator in the medium.

12. The method of claim 11 wherein the sample is added to a thin film containing the medium using a device having a self-supporting, waterproof substrate, a foam spacer and a transparent cover sheet, wherein a culture media is coated on the waterproof substrate and then dried.

13. A device to detect and enumerate Enterobacteriaceae in a sample consisting essentially of a coated self-supporting, waterproof substrate, a foam spacer and a transparent cover sheet, wherein a culture medium containing selected amounts of glucose, pH indicator and buffer to prevent colored indicator zones from excessively diffusing in the medium is coated on the self-supporting, waterproof substrate.